The microbiology of natural environments is still a relatively unexplored world; many hindrances prevent the knowledge of their actual biological diversity at bacterial level; among which the difficulty in culturing on plate media the vast majority of prokaryotic species. We sought to devise a polyphasic approach, comparing direct and culture-dependent methods, aimed at characterizing the bacterial communities rhizosphere and rhizoplane of herbaceous plants. The following tasks were pursued: 1) to define a procedure for the reliable and reproducible isolation of bacteria from the rhizospheric soil and from the rhizoplane of plants; 2) To compare the microbial communities of six spontaneous herbaceous plant species sharing the same natural habitat consisting in a ruderal vegetation site five years post-disturbance; the chosen plants are: Papaver rhoeas (Papaveraceae); Plantago lanceolata (Plantaginaceae) Daucus carota (Umbelliferae); Lotus corniculatus (Leguminosae); Taraxacum officinale (Compositae); and Lolium multiflorum (Graminaceae); 3) To analyze and count bacteria displaced from roots, using light and fluorescence microscopy visualizing: total cell counts (DAPI), live to dead cell ratio (Bac Light), and metabolically active cells (CTC+); 4) to count plate-culturable fractions of bacteria from rhizosphere and rhizoplane and assess percent culturability over the total cell counts obtained by microscopy; 5) to perform culture-independent DNA-based analyses by extracting total community DNA and PCR-amplifying 16S rDNA regions for ARDRA and DGGE comparisons. The results, comparing the outputs of: (1) computer assisted electrophoretic ARDRA fingerprinting, (2) community analysis by sequencing of 16S rDNA clone banks, (3) microscopical assessment of communities physiological status, will be discussed.

Comparative evaluation of rhizospheric and rhizoplanic bacterial communities of six spontaneous plant species sharing the same vegetation unit

VENDRAMIN, ELENA;ALBERGHINI, SARA;BASAGLIA, MARINA;SQUARTINI, ANDREA
2004

Abstract

The microbiology of natural environments is still a relatively unexplored world; many hindrances prevent the knowledge of their actual biological diversity at bacterial level; among which the difficulty in culturing on plate media the vast majority of prokaryotic species. We sought to devise a polyphasic approach, comparing direct and culture-dependent methods, aimed at characterizing the bacterial communities rhizosphere and rhizoplane of herbaceous plants. The following tasks were pursued: 1) to define a procedure for the reliable and reproducible isolation of bacteria from the rhizospheric soil and from the rhizoplane of plants; 2) To compare the microbial communities of six spontaneous herbaceous plant species sharing the same natural habitat consisting in a ruderal vegetation site five years post-disturbance; the chosen plants are: Papaver rhoeas (Papaveraceae); Plantago lanceolata (Plantaginaceae) Daucus carota (Umbelliferae); Lotus corniculatus (Leguminosae); Taraxacum officinale (Compositae); and Lolium multiflorum (Graminaceae); 3) To analyze and count bacteria displaced from roots, using light and fluorescence microscopy visualizing: total cell counts (DAPI), live to dead cell ratio (Bac Light), and metabolically active cells (CTC+); 4) to count plate-culturable fractions of bacteria from rhizosphere and rhizoplane and assess percent culturability over the total cell counts obtained by microscopy; 5) to perform culture-independent DNA-based analyses by extracting total community DNA and PCR-amplifying 16S rDNA regions for ARDRA and DGGE comparisons. The results, comparing the outputs of: (1) computer assisted electrophoretic ARDRA fingerprinting, (2) community analysis by sequencing of 16S rDNA clone banks, (3) microscopical assessment of communities physiological status, will be discussed.
2004
ISME X, 10th International Symposium on Microbial Ecology, Volume of Abstract
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11577/1335270
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